Pre-clinical characterization of GMP grade CCL21-gene modified dendritic cells for application in a phase I trial in Non-Small Cell Lung Cancer

Felicita Baratelli^*¹ , Hiroko Takedatsu^*¹ , Saswati Hazra¹ , Katherine Peebles¹ , Jie Luo¹ , Pam S Kurimoto¹ , Gang Zeng⁴ , Raj K Batra¹^,3 , Sherven Sharma¹^,3 , Steven M Dubinett¹^,2^,3 and Jay M Lee¹^,5

¹UCLA Lung Cancer Research Program of the Jonsson Comprehensive Cancer Center, Division of Pulmonary and Critical Care Medicine, Department of Medicine, Los Angeles, CA 90095, USA

²Department of Pathology and Laboratory Medicine, Geffen School of Medicine at UCLA, Los Angeles, CA, 90095, USA

³Molecular Medicine Laboratory, Veteran's Affairs Greater Los Angeles Healthcare System, Los Angeles, CA 90073, USA

⁴Department of Urology, Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA

⁵Division of Cardiothoracic Surgery, Department of Surgery, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA

author email corresponding author email* Contributed equally

Journal of Translational Medicine 2008, 6:38doi:10.1186/1479-5876-6-38


Published:	22 July 2008

Abstract

Background

Our previous studies have demonstrated that transduction of human dendritic cells (DC) with adenovirus encoding secondary lymphoid chemokine, CCL21, led to secretion of biologically active CCL21 without altering DC phenotype or viability. In addition, intratumoral injections of CCL21-transduced DC into established murine lung tumors resulted in complete regression and protective anti-tumor immunity. These results have provided the rationale to generate a clinical grade adenoviral vector encoding CCL-21 for ex vivo transduction of human DC in order to assess intratumoral administration in late stage human lung cancer.

Methods

In the current study, human monocyte-derived DC were differentiated by exposure to GM-CSF and IL-4 from cryopreserved mononuclear cells obtained from healthy volunteers. Transduction with clinical grade adenoviral vector encoding CCL21 (1167 viral particles per cell) resulted in secretion of CCL21 protein.

Results

CCL21 protein production from transduced DC was detected in supernatants (24–72 hours, 3.5–6.7 ng/4–5 × 10⁶cells). DC transduced with the clinical grade adenoviral vector were > 88% viable (n = 16), conserved their phenotype and maintained integral biological activities including dextran uptake, production of immunostimulatory cytokines/chemokines and antigen presentation. Furthermore, supernatant from CCL21-DC induced the chemotaxis of T2 cells in vitro.

Conclusion

Viable and biologically active clinical grade CCL21 gene-modified DC can be generated from cryopreserved PBMC.